Table I.
Determination of p53 protein amounts from different nuclear cell extracts.
| Different methods for determining p53 amounts | |||
|---|---|---|---|
| Cell source | ELISA | western | SPA |
| NPA 1 | 50+/−2 | 55+/−5 | 80+/−20 |
| 100+/−5 | 110+/−10 | 166+/−8 | |
| NPA 2 | 50+/−2 | 55+/−5 | 70+/−20 |
| 100+/−5 | 110+/−10 | 120+/−20 | |
| WRO | ND | 42+/−8 | 50+/−10 |
| ND | 80+/−10 | 73+/−7 | |
The amount of p53 (in pg) in the nuclear cell extracts was determined using 3 different methods- ELISA, western blots and SPA as described in Section 2. For the SPA, the values were determined from a standard curve with one ARO nuclear cell extract and then the counts from the other 3 extracts were compared to that standard curve. The equation for the SPA determined standard curve 240 minutes after adding the SPA beads was y=29.9x+1151 with an R2 value of 0.9926. These data are from one representative experiment out of a total of 6 similar experiments performed. ND- the p53 in the WRO cell extract was not detectable by ELISA.